Bayne C J, Menino J S, Hobbs D J, Barnes D W
Department of Zoology, Oregon State University, Corvallis 97331.
J Parasitol. 1994 Feb;80(1):29-35.
With the intent of providing a useful tool for studies on the cellular and molecular biology of Schistosoma mansoni, we have attempted to establish indefinitely proliferating cell lines. Primary (mother) sporocysts have served as sources of tissue fragments for initiation of primary cultures in complex media containing fetal bovine serum. Viability is maintained for several months, during which time there is differential survival of individual cell types. Cells that ultrastructurally resemble germinal cells are among the most persistent. Contractile responsiveness to serotonin and flagellar movement of flame cells are sustained for several weeks. Exposure to epidermal growth factor failed to induce tyrosine phosphorylation as detectable by western blot analysis. Incorporation of 5-bromo-2'-deoxyuridine into nucleic acid has been used as an indicator of the merit of experimental variables tested for their growth-promoting potential. Continuous proliferation remains an elusive goal, but coculture with host snail ganglia has yielded promising results. These primary cultures can be used to obtain useful information on parasite physiology. In light of our results, and of the varied lines of investigation that would be facilitated by such tools, further efforts to immortalize cell lines are warranted.
为了为曼氏血吸虫的细胞和分子生物学研究提供一个有用的工具,我们试图建立能无限增殖的细胞系。初代(母代)子胞蚴已作为组织碎片的来源,用于在含有胎牛血清的复杂培养基中启动原代培养。细胞活力可维持数月,在此期间不同类型的细胞有不同的存活率。超微结构上类似于生殖细胞的细胞是最持久的细胞类型之一。对血清素的收缩反应性和焰细胞的鞭毛运动可持续数周。通过蛋白质免疫印迹分析检测,暴露于表皮生长因子未能诱导酪氨酸磷酸化。将5-溴-2'-脱氧尿苷掺入核酸已被用作测试实验变量促进生长潜力优劣的指标。持续增殖仍然是一个难以实现的目标,但与宿主蜗牛神经节共培养已产生了有希望的结果。这些原代培养可用于获取有关寄生虫生理学的有用信息。鉴于我们的结果,以及此类工具将促进的各种研究方向,进一步努力使细胞系永生化是有必要的。